1. Detection of hemophilia A genetic variants using third-generation long-read sequencing.
- Author
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Ling, Xiaoting, Pan, Liqiu, Li, Linlin, Huang, Yunhua, Wang, Chenghan, Huang, Chaoyu, Long, Yan, Zhai, Ningneng, Xiao, Qingxing, Luo, Jiaqi, Tang, Rongheng, Meng, Li, and Huang, Yifang
- Subjects
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X-linked genetic disorders , *SINGLE nucleotide polymorphisms , *GENETIC testing , *GENETIC variation , *BLOOD coagulation factor VIII - Abstract
• Long-read sequencing shows great potential in detecting Hemophilia A with its unique advantages. • The detection of F8 gene based on Long-read sequencing provides a comprehensive characterization of pathogenic variants in a single step. • Long-read sequencing provides strong support for screening, genetic counseling, and prenatal diagnosis in carriers of Hemophilia A. Hemophilia A (HA) is an X-linked recessive genetic disorder caused by pathogenic variations of the factor VIII −encoding gene, F8 gene. Due to the large size and diverse types of variations in the F8 gene, causative mutations in F8 cannot be simultaneously detected in one step by traditional molecular analysis, and genetic molecular diagnosis and prenatal screening of HA still face significant difficulties and challenges in clinical practice. Therefore, we aimed to develop and validate an efficient, accurate, and time-saving method for the genetic detection of HA. A comprehensive analysis of hemophilia A (CAHEA) method based on long-range PCR and long-read sequencing (LRS) was used to detect F8 gene mutations in 14 clinical HA samples. The LRS results were compared with those of the conventional methods to evaluate the accuracy and sensitivity of the proposed approach. The CAHEA method successfully identified 14 F8 variants in all probands, including 3 small insertion deletions, 4 single nucleotide variants, and 7 intron 22 inversions in a "one-step" manner, of which 2 small deletions have not been reported previously. Moreover, this method provided an opportunity to analyze the mechanism of rearrangement and the pathogenicity of F8 variants. The LRS results were validated and found to be in 100% agreement with those obtained using the conventional method. Our proposed LRS-based F8 gene detection method is an accurate and reproducible genetic screening and diagnostic method with significant clinical value. It provides efficient, comprehensive, and accurate genetic screening and diagnostic services for individuals at high risk of HA as well as for premarital and prenatal populations. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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